The aims of this study were isolation and characterization of Pseudomonas spp. from fish meat collected from Sohag governorate, Egypt. A total 120 fish samples including frozen mackerel, frozen saurus, chilled Mugil cephalus and chilled Tilapia nilotica (30 of each) were collected from different shops and supermarkets in Sohag governorate. Pseudomonas spp. were isolated from 65% of the examined samples. The obtained data revealed that the highest count of Pseudomonas was in chilled Tilapia nilotica. The prevalence of Pseudomonas aeruginosa in frozen mackerel, frozen saurus, chilled Tilapia nilotica and chilled Mugil cephalus was 33.3%, 30%, 23.3% and 26.6% respectively. Furthermore, psychrotrophic count was performed and the results demonstrated that it was the highest in frozen mackerel followed by Tilapia nilotica and the Mugil cephalus showed the lowest count. Furthermore, the occurrence of oprL, phzM and toxA virulence genes was studied in some selected isolates by PCR. The findings showed that all the selected isolates possessed the virulence genes. This work showed contamination of fish samples with Pseudomonas spp., indicating the importance of applying hygienic measures during handling and storage of fish.